School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China.
Hangzhou Joinstar Biotechnology Co., Ltd., Hangzhou, 310000, China.
Talanta. 2023 Sep 1;262:124685. doi: 10.1016/j.talanta.2023.124685. Epub 2023 May 18.
Digital bio-detection has become one of the most appealing methods in recent years due to its excellent performance with ultra-sensitivity in detection of low-abundance targets. Traditional digital bio-detection needs the utilization of micro-chambers for physical isolation of targets, while the recently developed beads-based micro-chamber free one is attracting extensive attention, although there exist the disadvantages of overlaps between positive ("1") and negative ("0") signals as well as the decreased detection sensitivity in multiplexed mode. Here we propose a feasible and robust micro-chamber free digital bio-detection for multiplexed and ultrasensitive immunoassay based on encoded magnetic microbeads (EMMs) and tyramide signal amplification (TSA) strategy. An EMMs-based multiplexed platform is constructed by using a fluorescent encoding method, then a puissant signal amplification of positive events in TSA procedure is achieved via systematical revelation of key factors influences. For proof of concept, a three-plexed tumor markers detection is performed to evaluate our established platform. The detection sensitivity is comparable to the corresponding single-plexed assays and is also approximately 30-15,000 times improvement compared to the conventional suspension chip. Therefore, this multiplexed micro-chamber free digital bio-detection paves a promising way to be an ultrasensitive and powerful tool for clinical diagnosis.
数字生物检测近年来因其在超灵敏检测低丰度靶标方面的优异性能而成为最吸引人的方法之一。传统的数字生物检测需要利用微室进行目标的物理隔离，而最近开发的基于珠粒的无微室方法则引起了广泛关注，尽管存在正信号（“1”）和负信号（“0”）之间的重叠以及在多重检测模式下检测灵敏度降低的缺点。在这里，我们提出了一种基于编码磁性微珠（EMMs）和酪胺信号放大（TSA）策略的可行且稳健的无微室数字生物检测方法，用于多重和超灵敏免疫分析。通过荧光编码方法构建了基于 EMMs 的多重平台，然后通过系统揭示关键因素的影响，在 TSA 过程中实现了正事件的强大信号放大。为了验证概念，我们进行了三种肿瘤标志物的检测，以评估我们建立的平台。检测灵敏度可与相应的单重检测相媲美，与传统的悬浮芯片相比，灵敏度也提高了约 30-15000 倍。因此，这种无微室的多重数字生物检测为临床诊断提供了一种超灵敏、强大的工具。
